OXSENS · Intraprotein signalling in heme-based sensors: the oxygen sensor FixL
6РП — Действия „Мария Кюри“
- Период
- 2007-02-01 → 2008-06-30
- Финансиране от ЕС
- 160 487 €
- Участници
- 1
- Схема
- IIF
Линиите свързват координатора с партньорите. За проекти отпреди 2014 г. CORDIS не винаги дава точни координати. Тези точки са на ниво град или държава.
Накратко на български
Бактериалните сензори FixL и Dos се изследват, за да се разбере как протеините разпознават кислорода чрез специфични аминокиселини. Това помага да се обясни механизмът, по който тези молекули работят като превключватели, предаващи сигнали в клетката.
Кратко обяснение, генерирано от езиков модел по текста на CORDIS. Оригиналът е по-долу.
Резултати накратко
Final Activity Report Summary - OXSENS (Intraprotein signaling in heme-based sensors: the oxygen sensor FixL)
This project aimed at elucidating the mechanisms of intra-protein signalling and gaseous ligand discrimination in a heme-based gas sensor integrating molecular biology, biochemical, ultrafast spectroscopic, and computational techniques. As model systems, the bacterial oxygen sensors FixL and Dos were studied. These proteins contain a sensory PAS domain, which regulates the activity of an associated enzymatic domain via the binding/release of oxygen to the heme. In Dos, oxygen replaces an internal amino acid ligand, methionine 95, and this replacement initiates signalling. Dissociation of this internal ligand leads to an unusual slow rebinding (35 ps in addition to the generic 7-ps) phase. We have shown that the specific flexibility of the methionine side chain is at the origin of this behaviour and provided a model for the role of this flexibility in the early signalling events. In the homologous heme domain of FixL, oxygen does not replace an internal residue, but the strong rearrangements of an arginine residue that is in hydrogen bonding interaction with heme-bound oxygen is involved in signalling. We have shown that this arginine residue effectively and unusually 'cages' the heme-bound oxygen, a property that stabilizes the oxy-complex, and makes the protein act as a bistable switch. For both proteins, the roles of other residues in the heme vicinity, and the effect of the presence of the enzymatic domain on the heme domain have also been investigated.
Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз
Цел на проекта
This project aims at elucidating the mechanisms of intra-protein signalling and gaseous ligand discrimination in a heme-based gas sensor. As a model system, I propose to study the bacterial oxygen sensor FixL.This protein contains a sensory PAS domain, which regulates the activity of a histidine kinase domain via the binding/release of oxygen to the heme. Structures of the O2-bound and unliganded isolated heme domain are available and provide models for the start- and endpoints of the intra-domain signalling process.However, information on the intermediate pathway is lacking. In addition, the transmission to, and the influence of, the enzymatic domain, have not been taken into account. To address these issues, this proposal describes an approach integrating molecular biology, biochemical, ultrafast spectroscopic, and computational techniques.Based on structural information and simulations, substitutions of residues potentially involved in the signalling pathway are designed. The initial set of mutations concerns residues in the heme pocket and in the 'FG loop', which is located at the presumed interface between the constituents of the functional homodimer.The corresponding proteins will be over-expressed, as isolated sensor domain, as well as full-length proteins. Strategies are devised to address the challenging task of isolating stable and sufficient full-length proteins for spectroscopic analysis.To generate intermediates in the signalling pathway, the unique possibility to photocleave the heme-ligand bond will be exploited. Using ultrafast absorption and vibrational (Raman and infrared) spectroscopy, this allows the characterization of the electronic and structural properties of transient states and their dynamics, with femtosecond time resolution.Along with molecular dynamics modelling and with the perspective of ultrafast time resolved crystallography, pathways will be mapped and general features for intraprotein signal propagation proposed.
Оригинален текст от CORDIS (на английски).
Участници
- ECOLE POLYTECHNIQUE · PALAISEAUКоординаторНиво държаваГърция
Връзки
Данни: CORDIS, © Европейски съюз